Curcumin ameliorates oxidative stress-induced intestinal barrier injury and mitochondrial damage by promoting Parkin dependent mitophagy through AMPK-TFEB signal pathway
Curcumin ameliorates oxidative stress-induced intestinal barrier injury and mitochondrial damage by promoting Parkin dependent mitophagy through AMPK-TFEB signal pathway
复制标题
姜黄素通过 AMPK-TFEB 信号通路促进 Parkin 依赖性线粒体自噬，改善氧化应激诱导的肠道屏障损伤和线粒体损伤
DOI：
10.1016/j.freeradbiomed.2019.12.004
复制
发表时间：
2020-02-01
影响因子：
7.4
通讯作者：
Hu, Caihong
中科院分区：
文献类型：
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作者：
Cao, Shuting;Wang, Chunchun;Hu, Caihong
肠上皮细胞是保护机体免受有害抗原和病原体侵害的最关键屏障。氧化应激与肠屏障功能障碍有关。因此，迫切需要有效和安全的治疗方法来维持肠道氧化还原平衡。姜黄素通过抗氧化应激而具有广泛的有益生物学功能。然而，姜黄素是否能减轻氧化应激引起的肠道损伤和线粒体损伤仍不清楚。本研究发现，姜黄素能有效地改善过氧化氢（H2 O2）诱导的猪肠上皮细胞（IPEC-J2细胞）氧化应激、肠上皮屏障损伤和线粒体损伤，其机制与PTEN诱导的PINK 1-Parkin线粒体自噬有关。从机制上讲，帕金（一种线粒体自噬相关蛋白）的耗竭消除了姜黄素对H2 O2诱导的猪肠上皮细胞（IPEC-J2）的抗氧化应激、改善肠屏障和线粒体功能的保护作用。一致地，在用GFP-Parkin Delta UBL转染的细胞中未发现姜黄素的保护作用，所述GFP-Parkin Delta UBL编码没有泛素E3连接酶活性的突变体Parkin蛋白，表明姜黄素的保护作用需要Parkin的泛素E3连接酶。另一方面，我们还发现，当PRKAA 1在H2 O2处理的IPEC-J2细胞中耗尽时，姜黄素的保护功能减弱。免疫荧光和荧光素酶检测结果表明，姜黄素能显著增强H2 O2处理的IPEC-J2细胞核转位和转录因子EB（TFEB）的转录活性，且与AMPK抑制剂化合物C共同处理后，这种增强作用得到改善，表明姜黄素通过AMPK信号通路促进TFEB的转录。与体外数据一致，饮食姜黄素保护肠道屏障功能，改善氧化还原状态，减轻线粒体损伤，触发线粒体自噬，并影响AMPK-TFEB信号通路，在一个完善的猪氧化应激模型中，通过挑战敌草快。总之，这些结果揭示了姜黄素通过AMPK激活和随后的TFEB核转位诱导Parkin依赖性线粒体自噬来改善氧化应激，增强肠屏障功能和线粒体功能。
The gut epithelial is known as the most critical barrier for protection against harmful antigens and pathogens. Oxidative stress has been implicated in the dysfunction of the intestine barrier. Hence, effective and safe therapeutic approaches for maintaining intestinal redox balance are urgently needed. Curcumin has gained attention for its vast beneficial biological function via antioxidative stress. However, whether the curcumin can relief intestine damage and mitochondrial injury induced by oxidative stress is still unclear. In this study, we found that curcumin can effectively ameliorate hydrogen peroxide (H2O2)-induced oxidative stress, intestinal epithelial barrier injury and mitochondrial damage in porcine intestinal epithelial cells (IPEC-J2 cells) in a PTEN-induced putative kinase (PINK1)-Parkin mitophagy dependent way. Mechanistically, depletion of Parkin (a mitophagy related protein) abolished curcumin's protective action on anti-oxidative stress, improving intestinal barrier and mitochondrial function in porcine intestinal epithelial cells (IPEC-J2) induced by H2O2. Consistently, the protective effect of curcumin was not found in cells transfected with GFP-Parkin Delta UBL, which encodes a mutant Parkin protein without the ubiquitin E3 ligase activity, indicating that the ubiquitin E3 ligase of Parkin is required for curcumin's protective effects. On the other hand, we also found that the protective function of curcumin was diminished when PRKAA1 was depleted in IPEC-J2 cells treated with H2O2. Immunofluorescence and luciferase assay showed that curcumin dramatically enhanced nuclear translocation and transcriptional activity of transcription factor EB (TFEB) in IPEC-J2 cells treated with H2O2, and it was ameliorated by co-treated with compound C, an Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) inhibitor, which means curcumin promotes TFEB transcript via AMPK signal pathway. Consistent with in vitro data, dietary curcumin protected intestinal barrier function, improved redox status, alleviated mitochondrial damage, triggered mitophagy and influenced AMPK-TFEB signal pathway in a well-established pig oxidative stress model by challenging with diquat. Taken together, these results unveil that curcumin ameliorates oxidative stress, enhances intestinal barrier function and mitochondrial function via the induction of Parkin dependent mitophagy through AMPK activation and subsequent TFEB nuclear translocation.